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. 1997 Jun;40(6):731–738. doi: 10.1136/gut.40.6.731

In vitro evaluation of the role of antibodies against Helicobacter pylori in inhibiting adherence of the organism to gastric cells.

M Clyne 1, J Thomas 1, L Weaver 1, B Drumm 1
PMCID: PMC1027197  PMID: 9245926

Abstract

BACKGROUND: Once Helicobacter pylori infection is established, it is difficult to eradicate despite a persistent systemic and local immune response. It is not known whether immunisation can be used to prevent H pylori infection in humans. AIMS: To evaluate the effect of the human immune response on adherence of H pylori to gastric cells. METHODS: Human milk from a woman infected with H pylori and milk from a non-infected woman were each fractionated by chromatography on DEAE cellulose. Bacteria were incubated with either serum, human milk, human milk fractions, or secretory IgA before incubation with Kato III cells (cells from a gastric adenocarcinoma cell line). Bacterial adherence to the cells was assessed using flow cytometry. RESULTS: Serum from both the H pylori infected and non-infected women killed H pylori. This resulted from the action of complement as heating the serum to 56 degrees C for 30 minutes abolished the bactericidal activity. Immunoglobulin fractions from serum of both infected and non-infected women did not inhibit H pylori adherence to Kato III cells. Human milk from the woman infected with H pylori and from the non-infected woman inhibited binding of H pylori to Kato III cells by 50 to 70%. Secretory IgA isolated from human milk had minimal inhibitory effect on adherence and this was notably less than the inhibitory effect of whole human milk. CONCLUSIONS: Human milk inhibits adherence of H pylori to Kato III cells and this inhibition is independent of whether or not the donor is infected with H pylori. Secretory IgA has minimal inhibitory effect on H pylori adherence.

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  1. Andersson B., Porras O., Hanson L. A., Lagergård T., Svanborg-Edén C. Inhibition of attachment of Streptococcus pneumoniae and Haemophilus influenzae by human milk and receptor oligosaccharides. J Infect Dis. 1986 Feb;153(2):232–237. doi: 10.1093/infdis/153.2.232. [DOI] [PubMed] [Google Scholar]
  2. Batchelder M., Fox J. G., Hayward A., Yan L., Shames B., Murphy J. C., Palley L. Natural and experimental Helicobacter mustelae reinfection following successful antimicrobial eradication in ferrets. Helicobacter. 1996 Mar;1(1):34–42. doi: 10.1111/j.1523-5378.1996.tb00006.x. [DOI] [PubMed] [Google Scholar]
  3. Blaser M. J., Smith P. F., Kohler P. F. Susceptibility of Campylobacter isolates to the bactericidal activity of human serum. J Infect Dis. 1985 Feb;151(2):227–235. doi: 10.1093/infdis/151.2.227. [DOI] [PubMed] [Google Scholar]
  4. Borén T., Falk P., Roth K. A., Larson G., Normark S. Attachment of Helicobacter pylori to human gastric epithelium mediated by blood group antigens. Science. 1993 Dec 17;262(5141):1892–1895. doi: 10.1126/science.8018146. [DOI] [PubMed] [Google Scholar]
  5. Clyne M., Drumm B. Adherence of Helicobacter pylori to primary human gastrointestinal cells. Infect Immun. 1993 Oct;61(10):4051–4057. doi: 10.1128/iai.61.10.4051-4057.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Clyne M., Labigne A., Drumm B. Helicobacter pylori requires an acidic environment to survive in the presence of urea. Infect Immun. 1995 May;63(5):1669–1673. doi: 10.1128/iai.63.5.1669-1673.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Crabtree J. E., Taylor J. D., Wyatt J. I., Heatley R. V., Shallcross T. M., Tompkins D. S., Rathbone B. J. Mucosal IgA recognition of Helicobacter pylori 120 kDa protein, peptic ulceration, and gastric pathology. Lancet. 1991 Aug 10;338(8763):332–335. doi: 10.1016/0140-6736(91)90477-7. [DOI] [PubMed] [Google Scholar]
  8. Cravioto A., Tello A., Villafán H., Ruiz J., del Vedovo S., Neeser J. R. Inhibition of localized adhesion of enteropathogenic Escherichia coli to HEp-2 cells by immunoglobulin and oligosaccharide fractions of human colostrum and breast milk. J Infect Dis. 1991 Jun;163(6):1247–1255. doi: 10.1093/infdis/163.6.1247. [DOI] [PubMed] [Google Scholar]
  9. Czinn S. J., Cai A., Nedrud J. G. Protection of germ-free mice from infection by Helicobacter felis after active oral or passive IgA immunization. Vaccine. 1993;11(6):637–642. doi: 10.1016/0264-410x(93)90309-l. [DOI] [PubMed] [Google Scholar]
  10. Czinn S. J., Nedrud J. G. Oral immunization against Helicobacter pylori. Infect Immun. 1991 Jul;59(7):2359–2363. doi: 10.1128/iai.59.7.2359-2363.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Drumm B., Sherman P., Cutz E., Karmali M. Association of Campylobacter pylori on the gastric mucosa with antral gastritis in children. N Engl J Med. 1987 Jun 18;316(25):1557–1561. doi: 10.1056/NEJM198706183162501. [DOI] [PubMed] [Google Scholar]
  12. Drumm B., Sherman P. Long-term storage of Campylobacter pylori. J Clin Microbiol. 1989 Jul;27(7):1655–1656. doi: 10.1128/jcm.27.7.1655-1656.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eaton K. A., Brooks C. L., Morgan D. R., Krakowka S. Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun. 1991 Jul;59(7):2470–2475. doi: 10.1128/iai.59.7.2470-2475.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Ferrero R. L., Thiberge J. M., Huerre M., Labigne A. Recombinant antigens prepared from the urease subunits of Helicobacter spp.: evidence of protection in a mouse model of gastric infection. Infect Immun. 1994 Nov;62(11):4981–4989. doi: 10.1128/iai.62.11.4981-4989.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goodwin C. S., Armstrong J. A., Marshall B. J. Campylobacter pyloridis, gastritis, and peptic ulceration. J Clin Pathol. 1986 Apr;39(4):353–365. doi: 10.1136/jcp.39.4.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Graham D. Y. Treatment of peptic ulcers caused by Helicobacter pylori. N Engl J Med. 1993 Feb 4;328(5):349–350. doi: 10.1056/NEJM199302043280512. [DOI] [PubMed] [Google Scholar]
  17. Harboe N., Ingild A. Immunization, isolation of immunoglobulins, estimation of antibody titre. Scand J Immunol Suppl. 1973;1:161–164. doi: 10.1111/j.1365-3083.1973.tb03798.x. [DOI] [PubMed] [Google Scholar]
  18. Hayani K. C., Guerrero M. L., Morrow A. L., Gomez H. F., Winsor D. K., Ruiz-Palacios G. M., Cleary T. G. Concentration of milk secretory immunoglobulin A against Shigella virulence plasmid-associated antigens as a predictor of symptom status in Shigella-infected breast-fed infants. J Pediatr. 1992 Dec;121(6):852–856. doi: 10.1016/s0022-3476(05)80327-0. [DOI] [PubMed] [Google Scholar]
  19. Hentschel E., Brandstätter G., Dragosics B., Hirschl A. M., Nemec H., Schütze K., Taufer M., Wurzer H. Effect of ranitidine and amoxicillin plus metronidazole on the eradication of Helicobacter pylori and the recurrence of duodenal ulcer. N Engl J Med. 1993 Feb 4;328(5):308–312. doi: 10.1056/NEJM199302043280503. [DOI] [PubMed] [Google Scholar]
  20. Horiba N., Maekawa Y., Yamauchi Y., Ito M., Matsumoto T., Nakamura H. Complement activation by lipopolysaccharides purified from gram-negative bacteria isolated from infected root canals. Oral Surg Oral Med Oral Pathol. 1992 Nov;74(5):648–651. doi: 10.1016/0030-4220(92)90360-3. [DOI] [PubMed] [Google Scholar]
  21. Jensen E. T., Kharazmi A., Garred P., Kronborg G., Fomsgaard A., Mollnes T. E., Høiby N. Complement activation by Pseudomonas aeruginosa biofilms. Microb Pathog. 1993 Nov;15(5):377–388. doi: 10.1006/mpat.1993.1087. [DOI] [PubMed] [Google Scholar]
  22. Jones G. W., Freter R. Adhesive properties of Vibrio cholerae: nature of the interaction with isolated rabbit brush border membranes and human erythrocytes. Infect Immun. 1976 Jul;14(1):240–245. doi: 10.1128/iai.14.1.240-245.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Korhonen H., Syväoja E. L., Ahola-Luttila H., Sivelä S., Kopola S., Husu J., Kosunen T. U. Bactericidal effect of bovine normal and immune serum, colostrum and milk against Helicobacter pylori. J Appl Bacteriol. 1995 Jun;78(6):655–662. doi: 10.1111/j.1365-2672.1995.tb03112.x. [DOI] [PubMed] [Google Scholar]
  24. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  25. Lee A., Chen M. Successful immunization against gastric infection with Helicobacter species: use of a cholera toxin B-subunit-whole-cell vaccine. Infect Immun. 1994 Aug;62(8):3594–3597. doi: 10.1128/iai.62.8.3594-3597.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Marshall B. J., Warren J. R. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet. 1984 Jun 16;1(8390):1311–1315. doi: 10.1016/s0140-6736(84)91816-6. [DOI] [PubMed] [Google Scholar]
  27. Michetti P., Corthésy-Theulaz I., Davin C., Haas R., Vaney A. C., Heitz M., Bille J., Kraehenbuhl J. P., Saraga E., Blum A. L. Immunization of BALB/c mice against Helicobacter felis infection with Helicobacter pylori urease. Gastroenterology. 1994 Oct;107(4):1002–1011. doi: 10.1016/0016-5085(94)90224-0. [DOI] [PubMed] [Google Scholar]
  28. Mitchell H. M., Li Y. Y., Hu P. J., Liu Q., Chen M., Du G. G., Wang Z. J., Lee A., Hazell S. L. Epidemiology of Helicobacter pylori in southern China: identification of early childhood as the critical period for acquisition. J Infect Dis. 1992 Jul;166(1):149–153. doi: 10.1093/infdis/166.1.149. [DOI] [PubMed] [Google Scholar]
  29. Pappo J., Thomas W. D., Jr, Kabok Z., Taylor N. S., Murphy J. C., Fox J. G. Effect of oral immunization with recombinant urease on murine Helicobacter felis gastritis. Infect Immun. 1995 Apr;63(4):1246–1252. doi: 10.1128/iai.63.4.1246-1252.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Rathbone B. J., Wyatt J. I., Worsley B. W., Shires S. E., Trejdosiewicz L. K., Heatley R. V., Losowsky M. S. Systemic and local antibody responses to gastric Campylobacter pyloridis in non-ulcer dyspepsia. Gut. 1986 Jun;27(6):642–647. doi: 10.1136/gut.27.6.642. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rautelin H., Blomberg B., Fredlund H., Järnerot G., Danielsson D. Incidence of Helicobacter pylori strains activating neutrophils in patients with peptic ulcer disease. Gut. 1993 May;34(5):599–603. doi: 10.1136/gut.34.5.599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Ruiz-Palacios G. M., Calva J. J., Pickering L. K., Lopez-Vidal Y., Volkow P., Pezzarossi H., West M. S. Protection of breast-fed infants against Campylobacter diarrhea by antibodies in human milk. J Pediatr. 1990 May;116(5):707–713. doi: 10.1016/s0022-3476(05)82652-6. [DOI] [PubMed] [Google Scholar]
  33. Schroten H., Hanisch F. G., Plogmann R., Hacker J., Uhlenbruck G., Nobis-Bosch R., Wahn V. Inhibition of adhesion of S-fimbriated Escherichia coli to buccal epithelial cells by human milk fat globule membrane components: a novel aspect of the protective function of mucins in the nonimmunoglobulin fraction. Infect Immun. 1992 Jul;60(7):2893–2899. doi: 10.1128/iai.60.7.2893-2899.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Strömqvist M., Falk P., Bergström S., Hansson L., Lönnerdal B., Normark S., Hernell O. Human milk kappa-casein and inhibition of Helicobacter pylori adhesion to human gastric mucosa. J Pediatr Gastroenterol Nutr. 1995 Oct;21(3):288–296. doi: 10.1097/00005176-199510000-00006. [DOI] [PubMed] [Google Scholar]
  35. Thomas J. E., Austin S., Dale A., McClean P., Harding M., Coward W. A., Weaver L. T. Protection by human milk IgA against Helicobacter pylori infection in infancy. Lancet. 1993 Jul 10;342(8863):121–121. doi: 10.1016/0140-6736(93)91327-i. [DOI] [PubMed] [Google Scholar]
  36. Williams R. C., Gibbons R. J. Inhibition of bacterial adherence by secretory immunoglobulin A: a mechanism of antigen disposal. Science. 1972 Aug 25;177(4050):697–699. doi: 10.1126/science.177.4050.697. [DOI] [PubMed] [Google Scholar]
  37. Wyatt J. I., Rathbone B. J., Dixon M. F., Heatley R. V. Campylobacter pyloridis and acid induced gastric metaplasia in the pathogenesis of duodenitis. J Clin Pathol. 1987 Aug;40(8):841–848. doi: 10.1136/jcp.40.8.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Yolken R. H., Peterson J. A., Vonderfecht S. L., Fouts E. T., Midthun K., Newburg D. S. Human milk mucin inhibits rotavirus replication and prevents experimental gastroenteritis. J Clin Invest. 1992 Nov;90(5):1984–1991. doi: 10.1172/JCI116078. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Zhang T., Li E., Stanley S. L., Jr Oral immunization with the dodecapeptide repeat of the serine-rich Entamoeba histolytica protein (SREHP) fused to the cholera toxin B subunit induces a mucosal and systemic anti-SREHP antibody response. Infect Immun. 1995 Apr;63(4):1349–1355. doi: 10.1128/iai.63.4.1349-1355.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]

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